【摘要】 目的 :探讨缺血再灌注 (缺氧再给氧 )对内皮细胞免疫功能的影响及川芎醇的干预效果。方法 :用人脐静脉内皮细胞株ECV30 4缺氧再给氧模型模拟体内缺血再灌注对血管内皮细胞的损伤 ,并用川芎醇干预其缺氧再给氧过程。间接免疫荧光染色显示胞内核因子 κΒ(ΝF κΒ) ,流式细胞术检测细胞表面HLA ABC ,HLA DR ,CD86和CD5 4 ,并将内皮细胞与同种异体淋巴细胞混合培养 (MELR) ,检测淋巴细胞增殖程度、IL 2的产生及凋亡淋巴细胞百分率。结果 :缺氧再给氧使ECV30 4细胞变圆、收缩、脱落 ,胞浆NF κB表达增高 ,且由胞浆阳性为主转为胞核阳性为主 ,细胞表面HLA ABC ,HLA DR ,CD86增高 ,CD5 4无显著变化 ,MELR中3 H TdR掺入量、IL 2生成量显著增多 ,凋亡淋巴细胞百分率降低。川芎醇干预后ECV30 4细胞形态基本恢复正常 ,NF κB表达未下调 ,但核阳性细胞百分率降低 ,以核浆阳性为主 ,其他多项检测指标的改变得以逆转。结论 :缺氧再给氧影响了ECV30 4细胞几种重要免疫相关分子的表达 ,川芎醇可发挥拮抗作用 ,使内皮细胞保持相对正常的免疫学功能更多还原

【Abstract】 Objective:To explore the influence of ischemia/reperfusion (anoxia/reoxygenation)[FK(W16*2。142mmZQ1mm]on immunofunction of endothelial cells(ECs) and effect of intervention with 2-hydroxymethyl-3,5,6-trimethylpyrazine on it.Methods:Model of ECs induced by anoxia/reoxygenation was established to mimic ECs ischemia/reperfusion injury in vivo with human umbilical vein endothelial cell line ECV304.Then 2-hydroxymethyl-3,5,6-trimethylpyraxine was used to intervene the anoxia/reoxygenation process.Nuclear transcriptional factor-κB(NF-κB) was exhibited by fluorescent staining,HLA-ABC,HLA-DR,CD86 and CD54 were detected by flow cytometry.Mixed endothelial cell-lymphocyte reaction(MELR) was conducted to examine the proliferation of lymphocyte,production of IL-2 and percentage of apoptotic lymphocyte.Results:Anoxia/reoxygenation made the ECV304 cell became round,shrunk and abscised,with increased plasma NF-κB,and changed from positive cytoplasm to positive nucleus.HLA-ABC,HLA-DR and CD86 on surface of cells increased but CD54 showed unchanged.MELR showed the incorporation of ~3H-TdR and production of IL-2 increased significantly and the percentage of apoptotic lymphocyte decreased.After 2-hydroxymethyl-3,5,6-trimethylpyrazine intervention,the ECV304 cell shapes recovered,NF-κB expression didn’t down-regulated,but the percentage of positive cells decreased,changes to positve dominant.Besides,reversal changes were shown in other parameters.Conclusion:Anoxia/reoxygenation influences some important immune related molecules in ECV304 cells.2-hydroxymethyl-3,5,6-trimethylpyrazine could antagonize these influences to maintain the immune function of endothelial cells in a relative normal manner.更多还原